MetaData in Bioinformatics Research


Accessing (Meta)Data from Gene Expression Omnibus (GEO) using R


GEOquery

GEO database at the National Center for Biotechnology Information and European Bioinformatics Institute (EBI) ArrayExpress follow the MIAME format for data storage. Minimum Information About a Microarray Experiment (MIAME) is a set of guidelines developed by the Microarray Gene Expression Data (MGED). It been adopted by many journals to support an independent evaluation of results.

Among the several R packages that can be used to access its data, GEOquery is a simple and widely used one.
To start, we will need

if (!require("BiocManager", quietly = TRUE)) install.packages("BiocManager")
BiocManager::install("GEOquery", force = TRUE)
library(GEOquery)
library(tidyverse)
library(pryr)

Data from GEO is access by a GEO Accession ID

Datasets: GDS[0-9] :

  • Curated set of GEO sample data
  • Samples in a given GDS share the same Platform
  • Methodology is assumed to be consistent

Sample: GSM[0-9]

  • Records information about a
    • Single Sample
    • Cell line / tissue
    • References a single Platform
    • May be in multiple Series

Data from GEO is access by a GEO Accession ID

Platform | GPL[0-9]

  • Provides a summary description of the experimental Platform. i.e. Microarray, Sequencer
  • ID is not limited to a single sample or submitter

Series: GSE[0-9]

  • Similar to datasets, it defines a set of related Samples
  • Can have data from multiple platform
  • Available in two formats:
    • ExpressionSet and Biobase object
  • Often, multiple series are combined to create a Superseries.

getGEO

The main user-level function in the GEOquery package is “getGEO”

args(getGEO)
## function (GEO = NULL, filename = NULL, destdir = tempdir(), GSElimits = NULL, 
##     GSEMatrix = TRUE, AnnotGPL = FALSE, getGPL = TRUE, parseCharacteristics = TRUE) 
## NULL

To download the data to a specific folder, we will need to create one. We could also provide an exisiting location.

# To store the data to specified folder, we need the the Dataset,
dir.create("GeoData")

Downloading a GDS object

We will access the data submitted in the following article,
“Recurrent somatic mutations of PTPN1 in primary mediastinal B cell lymphoma and Hodgkin lymphoma”. Nat Genet 2014 Apr;46(4):329-35

gds <- GEOquery::getGEO(GEO = "GDS5288", destdir = "GeoData")

Accesor functions

Meta() returns a list of object metadata

GEOquery::Meta(gds)[c("title", "description", "ref", "platform", "platform_technology_type",
    "sample_organism", "sample_id", "sample_type")]
## $title
## [1] "PTPN1 depletion effect on Hodgkin lymphoma derived cell line"
## 
## $description
## [1] "Analysis of KM-H2 Hodgkin lymphoma derived cells depleted for PTPN1. PTPN1 encodes PTP1B, a non-receptor-type member of the superfamily of protein tyrosine phosphatases. Results provide insight into the role of PTPN1 mutations in lymphomagenesis."
## [2] "PTPN1 depletion"                                                                                                                                                                                                                                       
## [3] "control"                                                                                                                                                                                                                                               
## 
## $ref
## [1] "Nucleic Acids Res. 2005 Jan 1;33 Database Issue:D562-6"
## 
## $platform
## [1] "GPL570"
## 
## $platform_technology_type
## [1] "in situ oligonucleotide"
## 
## $sample_organism
## [1] "Homo sapiens"
## 
## $sample_id
## [1] "GSM1308977,GSM1308978" "GSM1308975,GSM1308976"
## 
## $sample_type
## [1] "RNA"

Accesor functions

Columns() returns a data.frame of column descriptors. This includes information about each of the samples in the data.

GEOquery::Columns(gds)
##       sample        protocol
## 1 GSM1308977 PTPN1 depletion
## 2 GSM1308978 PTPN1 depletion
## 3 GSM1308975         control
## 4 GSM1308976         control
##                                                                                          description
## 1 Value for GSM1308977: KM-H2 PTPN1 Knockdown, technical rep1; src: KM-H2 with PTPN1 siRNA knockdown
## 2 Value for GSM1308978: KM-H2 PTPN1 Knockdown, technical rep2; src: KM-H2 with PTPN1 siRNA knockdown
## 3                Value for GSM1308975: KM-H2 Non-silencing, technical rep1; src: KM-H2 non-silencing
## 4                Value for GSM1308976: KM-H2 Non-silencing, technical rep2; src: KM-H2 non-silencing

Accesor functions

Table() returns a data.frame of gene expression data. The columns include gene IDs and expression values by Sample.

GEOquery::Table(gds)[1:5, ]
##      ID_REF IDENTIFIER GSM1308977 GSM1308978 GSM1308975 GSM1308976
## 1 1007_s_at    MIR4640    8.82235    8.86388    8.58477    8.72635
## 2   1053_at       RFC2   10.03960    9.81700   10.08130    9.80014
## 3    117_at      HSPA6    5.98696    6.20082    6.18230    6.70773
## 4    121_at       PAX8    7.74991    8.02246    7.94855    8.20815
## 5 1255_g_at     GUCA1A    4.10924    3.83838    3.88709    4.00901

The GDS object

pryr::object_size(gds)
## 7.86 MB
class(gds)
## [1] "GDS"
## attr(,"package")
## [1] "GEOquery"
isS4(gds)
## [1] TRUE

GSM - Sample

There are similar workflows for Sample, Series and Platform.

gsm <- getGEO("GSM1308977", destdir = "GeoData")
Meta(gsm)
## $channel_count
## [1] "1"
## 
## $characteristics_ch1
## [1] "tissue: Hodgkin lymphoma" "cell line: KM-H2"        
## 
## $contact_address
## [1] "675 West 10th Avenue"
## 
## $contact_city
## [1] "Vancouver"
## 
## $contact_country
## [1] "Canada"
## 
## $contact_department
## [1] "Pathology"
## 
## $contact_email
## [1] "csteidl@bccancer.bc.ca"
## 
## $contact_institute
## [1] "BC Cancer Agency"
## 
## $contact_laboratory
## [1] "Steidl lab"
## 
## $contact_name
## [1] "Christian,,Steidl"
## 
## $contact_phone
## [1] "1-604-675-8046"
## 
## $contact_state
## [1] "BC"
## 
## $`contact_zip/postal_code`
## [1] "V5Z 1L3"
## 
## $data_processing
## [1] "The R package aroma.affymetrix (v2.9.0) from Bioconductor was used to generate RMA expression and perform QC."
## 
## $data_row_count
## [1] "54675"
## 
## $extract_protocol_ch1
## [1] "Qiagen Allprep extraction columns"
## 
## $geo_accession
## [1] "GSM1308977"
## 
## $growth_protocol_ch1
## [1] "Cell lines: Standard conditions (www.dsmz.de)"
## 
## $hyb_protocol
## [1] "Following fragmentation, cRNA were hybridized O/N at 45C on Affymetrix GeneChip HG 2.0 Plus Arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station 450."
## 
## $label_ch1
## [1] "biotin"
## 
## $label_protocol_ch1
## [1] "Biotinylated cRNA were prepared according to the standard Affymetrix protocol (2-cycle amplification)"
## 
## $last_update_date
## [1] "Jan 17 2014"
## 
## $molecule_ch1
## [1] "total RNA"
## 
## $organism_ch1
## [1] "Homo sapiens"
## 
## $platform_id
## [1] "GPL570"
## 
## $scan_protocol
## [1] "GeneChips were scanned using Affymetrix GeneChip Scanner"
## 
## $series_id
## [1] "GSE54157"
## 
## $source_name_ch1
## [1] "KM-H2 with PTPN1 siRNA knockdown"
## 
## $status
## [1] "Public on Jan 17 2014"
## 
## $submission_date
## [1] "Jan 16 2014"
## 
## $supplementary_file
## [1] "ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM1308nnn/GSM1308977/suppl/GSM1308977_KMH2-PTPN1KD1.CEL.gz"
## 
## $taxid_ch1
## [1] "9606"
## 
## $title
## [1] "KM-H2 PTPN1 Knockdown, technical rep1"
## 
## $type
## [1] "RNA"

GSM - Sample

Similar accessor functions work

Columns(gsm)
##   Column                Description
## 1 ID_REF                           
## 2  VALUE log2 RMA normalized signal
Table(gsm)
##                            ID_REF     VALUE
## 1                  AFFX-BioB-5_at  8.480296
## 2                  AFFX-BioB-M_at  9.087702
## 3                  AFFX-BioB-3_at  8.707758
## 4                  AFFX-BioC-5_at  9.845029
## 5                  AFFX-BioC-3_at 10.415414
## 6                 AFFX-BioDn-5_at 11.051690
## 7                 AFFX-BioDn-3_at 12.035703
## 8                  AFFX-CreX-5_at 13.612359
## 9                  AFFX-CreX-3_at 13.632270
## 10                 AFFX-DapX-5_at  7.975419
## 11                 AFFX-DapX-M_at 10.315585
## 12                 AFFX-DapX-3_at 10.890056
## 13                 AFFX-LysX-5_at  6.740159
## 14                 AFFX-LysX-M_at  7.525384
## 15                 AFFX-LysX-3_at  7.924963
## 16                 AFFX-PheX-5_at  7.571823
## 17                 AFFX-PheX-M_at  7.816444
## 18                 AFFX-PheX-3_at  7.906589
## 19                 AFFX-ThrX-5_at  6.523586
## 20                 AFFX-ThrX-M_at  7.921625
## 21                 AFFX-ThrX-3_at  9.298156
## 22                AFFX-TrpnX-5_at  3.860871
## 23                AFFX-TrpnX-M_at  3.432896
## 24                AFFX-TrpnX-3_at  3.375616
## 25           AFFX-r2-Ec-bioB-5_at  8.921402
## 26           AFFX-r2-Ec-bioB-M_at  9.382877
## 27           AFFX-r2-Ec-bioB-3_at  9.190695
## 28           AFFX-r2-Ec-bioC-5_at 10.524649
## 29           AFFX-r2-Ec-bioC-3_at 10.951632
## 30           AFFX-r2-Ec-bioD-5_at 12.445226
## 31           AFFX-r2-Ec-bioD-3_at 12.661747
## 32            AFFX-r2-P1-cre-5_at 14.112683
## 33            AFFX-r2-P1-cre-3_at 14.307971
## 34            AFFX-r2-Bs-dap-5_at  8.819005
## 35            AFFX-r2-Bs-dap-M_at 10.671276
## 36            AFFX-r2-Bs-dap-3_at 11.319084
## 37            AFFX-r2-Bs-lys-5_at  7.539878
## 38            AFFX-r2-Bs-lys-M_at  7.334497
## 39            AFFX-r2-Bs-lys-3_at  7.483875
## 40            AFFX-r2-Bs-phe-5_at  7.930399
## 41            AFFX-r2-Bs-phe-M_at  8.124227
## 42            AFFX-r2-Bs-phe-3_at  8.738725
## 43          AFFX-r2-Bs-thr-3_s_at  9.716796
## 44          AFFX-r2-Bs-thr-M_s_at  8.520607
## 45          AFFX-r2-Bs-thr-5_s_at  6.876051
## 46     AFFX-HUMISGF3A/M97935_5_at  6.641523
## 47    AFFX-HUMISGF3A/M97935_MA_at  7.896011
## 48    AFFX-HUMISGF3A/M97935_MB_at  8.464774
## 49     AFFX-HUMISGF3A/M97935_3_at 10.621345
## 50        AFFX-HUMRGE/M10098_5_at 13.464086
## 51        AFFX-HUMRGE/M10098_M_at 12.116453
## 52        AFFX-HUMRGE/M10098_3_at 11.691937
## 53      AFFX-HUMGAPDH/M33197_5_at 13.722883
## 54      AFFX-HUMGAPDH/M33197_M_at 13.912943
## 55      AFFX-HUMGAPDH/M33197_3_at 14.015626
## 56        AFFX-HSAC07/X00351_5_at 11.537511
## 57        AFFX-HSAC07/X00351_M_at 12.292272
## 58        AFFX-HSAC07/X00351_3_at 12.864160
## 59               AFFX-M27830_5_at 12.512481
## 60               AFFX-M27830_M_at  9.820761
## 61               AFFX-M27830_3_at  5.920335
## 62                AFFX-hum_alu_at 14.461802
## 63                      1007_s_at  8.822346
## 64                        1053_at 10.039576
## 65                         117_at  5.986958
## 66                         121_at  7.749908
## 67                      1255_g_at  4.109242
## 68                        1294_at  5.745199
## 69                        1316_at  6.573133
## 70                        1320_at  4.426695
## 71                      1405_i_at 11.308156
## 72                        1431_at  3.689785
## 73                        1438_at  4.754326
## 74                        1487_at  8.317358
## 75                      1494_f_at  5.209210
## 76                      1598_g_at  6.691646
## 77                      160020_at  7.128541
## 78                        1729_at  8.881501
## 79                        1773_at  6.047666
## 80                         177_at  5.230498
## 81                         179_at  7.273010
## 82                        1861_at  8.096116
## 83                    200000_s_at  9.659203
## 84                      200001_at  8.581477
## 85                      200002_at 13.783214
## 86                    200003_s_at 14.115319
## 87                      200004_at 12.904310
## 88                      200005_at 11.582475
## 89                      200006_at 13.384057
## 90                      200007_at 12.242126
## 91                    200008_s_at 10.043928
## 92                      200009_at 11.984045
## 93                      200010_at 13.698669
## 94                    200011_s_at  9.904530
## 95                    200012_x_at 13.968094
## 96                      200013_at 13.549348
## 97                    200014_s_at 12.719560
## 98                    200015_s_at 11.288356
## 99                    200016_x_at 14.117193
## 100                     200017_at 13.370158
## 101                     200018_at 14.249267
## 102                   200019_s_at 13.497728
## 103                     200020_at 11.500300
## 104                     200021_at 11.406532
## 105                     200022_at 13.603749
## 106                   200023_s_at 13.236288
## 107                     200024_at 13.780158
## 108                   200025_s_at 14.168538
## 109                     200026_at 12.985872
## 110                     200027_at 11.849107
## 111                   200028_s_at 11.509623
## 112                     200029_at 13.872857
## 113                   200030_s_at 13.597197
## 114                   200031_s_at 14.258649
## 115                   200032_s_at 14.109774
## 116                     200033_at 12.705432
## 117                   200034_s_at 13.942404
## 118                     200035_at  9.026003
## 119                   200036_s_at 13.457357
## 120                   200037_s_at 12.475472
## 121                   200038_s_at 13.791891
## 122                   200039_s_at 11.331810
## 123                     200040_at 12.193877
## 124                   200041_s_at  9.374694
## 125                     200042_at 10.073229
## 126                     200043_at 13.372447
## 127                     200044_at 12.839004
## 128                     200045_at 10.424759
## 129                     200046_at 11.254728
## 130                   200047_s_at 12.327286
## 131                   200048_s_at 12.512174
## 132                     200049_at  7.870194
## 133                     200050_at 10.355735
## 134                     200051_at  9.440892
## 135                   200052_s_at 12.083900
## 136                     200053_at  9.803635
## 137                     200054_at  9.146030
## 138                     200055_at 11.200931
## 139                   200056_s_at 10.447312
## 140                   200057_s_at 12.529187
## 141                   200058_s_at 11.616815
## 142                   200059_s_at 12.536496
## 143                   200060_s_at 10.549858
## 144                   200061_s_at 14.439457
## 145                   200062_s_at 14.013908
## 146                   200063_s_at 13.947421
## 147                     200064_at 12.851867
## 148                   200065_s_at 12.040052
## 149                     200066_at 10.269893
## 150                   200067_x_at  9.915797
## 151                   200068_s_at 12.924533
## 152                     200069_at  9.546988
## 153                     200070_at  9.223257
## 154                     200071_at 10.616998
## 155                   200072_s_at 12.324818
## 156                   200073_s_at 12.424807
## 157                   200074_s_at 13.313639
## 158                   200075_s_at 11.413457
## 159                   200076_s_at  9.907296
## 160                   200077_s_at 13.717573
## 161                   200078_s_at 10.987142
## 162                   200079_s_at 11.861610
## 163                   200080_s_at 13.585725
## 164                   200081_s_at 14.332478
## 165                   200082_s_at 14.298171
## 166                     200083_at 10.897697
## 167                     200084_at 11.732397
## 168                   200085_s_at 11.855858
## 169                   200086_s_at 13.202663
## 170                   200087_s_at 12.430655
## 171                   200088_x_at 14.146580
## 172                   200089_s_at 13.826362
## 173                     200090_at 10.419256
## 174                   200091_s_at 13.425585
## 175                   200092_s_at 14.255481
## 176                   200093_s_at 12.833255
## 177                   200094_s_at 12.588866
## 178                   200095_x_at 14.019688
## 179                   200096_s_at 11.241320
## 180                   200097_s_at 11.874733
## 181                   200098_s_at 12.119228
## 182                   200099_s_at 14.314440
## 183                   200593_s_at 11.935262
## 184                   200594_x_at 12.937516
## 185                   200595_s_at 10.832740
## 186                   200596_s_at 10.761343
## 187                     200597_at  8.075234
## 188                   200598_s_at 12.931752
## 189                   200599_s_at 12.578343
## 190                     200600_at 10.435159
## 191                     200601_at  7.429719
## 192                     200602_at  2.480872
## 193                     200603_at 10.572433
## 194                   200604_s_at  6.945951
## 195                   200605_s_at 10.608057
## 196                     200606_at  3.436413
## 197                   200607_s_at 10.789473
## 198                   200608_s_at 12.474601
## 199                   200609_s_at 10.209768
## 200                   200610_s_at 12.668208
## 201                   200611_s_at  9.134033
## 202                   200612_s_at  8.912708
## 203                     200613_at  9.203429
## 204                     200614_at 12.480836
## 205                   200615_s_at  7.555597
## 206                   200616_s_at 11.747769
## 207                     200617_at  8.814205
## 208                     200618_at  9.465431
## 209                     200619_at 11.207956
## 210                     200620_at 10.552594
## 211                     200621_at 10.434185
## 212                   200622_x_at  9.089468
## 213                   200623_s_at  8.500047
## 214                   200624_s_at 11.193746
## 215                   200625_s_at 11.456308
## 216                   200626_s_at 11.422671
## 217                     200627_at 13.388816
## 218                   200628_s_at  8.581378
## 219                     200629_at  9.777115
## 220                   200630_x_at 12.934858
## 221                   200631_s_at 12.658453
## 222                   200632_s_at  5.928187
## 223                     200633_at 14.336119
## 224                     200634_at 11.855378
## 225                   200635_s_at  6.823077
## 226                   200636_s_at  7.702754
## 227                   200637_s_at  5.588088
## 228                   200638_s_at 11.668855
## 229                   200639_s_at 10.136258
## 230                     200640_at 10.259756
## 231                   200641_s_at 10.076624
## 232                     200642_at 13.136612
## 233                     200643_at  8.087903
## 234                     200644_at 10.143557
## 235                     200645_at  9.898149
## 236                   200646_s_at  5.042591
## 237                   200647_x_at 11.035589
## 238                   200648_s_at  5.639544
## 239                     200649_at  6.621255
## 240                   200650_s_at 13.736745
## 241                     200651_at 13.999929
## 242                     200652_at 12.210452
## 243                   200653_s_at 11.863416
## 244                     200654_at  8.989660
## 245                   200655_s_at 12.615099
## 246                   200656_s_at  9.647208
## 247                     200657_at 13.241386
## 248                   200658_s_at 11.757978
## 249                   200659_s_at  9.389960
## 250                     200660_at  8.505558
## 251                     200661_at  8.350532
## 252                   200662_s_at 11.506318
## 253                     200663_at 11.925663
## 254                   200664_s_at  8.329539
## 255                   200665_s_at 10.717311
## 256                   200666_s_at  9.098935
## 257                     200667_at 11.991216
## 258                   200668_s_at 13.154673
## 259                   200669_s_at 12.266114
## 260                     200670_at  8.580168
## 261                   200671_s_at  6.332543
## 262                   200672_x_at  7.652471
## 263                     200673_at 11.553467
## 264                   200674_s_at 14.329744
## 265                     200675_at 11.332579
## 266                   200676_s_at  8.308081
## 267                     200677_at 10.443614
## 268                   200678_x_at  9.251905
## 269                   200679_x_at 12.419026
## 270                   200680_x_at 13.261962
## 271                     200681_at 11.334599
## 272                   200682_s_at 12.756956
## 273                   200683_s_at 10.859556
## 274                   200684_s_at  9.560726
## 275                     200685_at  7.734057
## 276                   200686_s_at 10.770569
## 277                   200687_s_at 11.009125
## 278                     200688_at  7.920688
## 279                   200689_x_at 13.960051
## 280                     200690_at  9.174451
## 281                   200691_s_at 12.003473
## 282                   200692_s_at 11.325010
## 283                     200693_at 13.148701
## 284                   200694_s_at 11.373793
## 285                     200695_at  7.372440
## 286                   200696_s_at  5.120492
## 287                     200697_at 10.036311
## 288                     200698_at  9.816964
## 289                     200699_at  9.066497
## 290                   200700_s_at 10.259213
## 291                     200701_at 10.683507
## 292                   200702_s_at 10.536843
## 293                     200703_at 12.744729
## 294                     200704_at 11.807016
## 295                   200705_s_at 13.913018
## 296                   200706_s_at 11.060022
## 297                     200707_at  6.741474
## 298                     200708_at 10.733748
## 299                     200709_at  8.627798
## 300                     200710_at  7.922733
## 301                   200711_s_at 10.625070
## 302                   200712_s_at  8.493910
## 303                   200713_s_at 11.093338
## 304                   200714_x_at  8.637788
## 305                   200715_x_at 13.187107
## 306                   200716_x_at 14.314744
## 307                   200717_x_at 14.105230
## 308                   200718_s_at 13.409335
## 309                     200719_at  8.746037
## 310                   200720_s_at  7.500802
## 311                   200721_s_at  7.177285
## 312                   200722_s_at  8.480612
## 313                   200723_s_at 11.357052
## 314                     200724_at  5.127221
## 315                   200725_x_at 13.962266
## 316                     200726_at 12.934477
## 317                   200727_s_at  9.788947
## 318                     200728_at 11.951280
## 319                   200729_s_at 11.794086
## 320                   200730_s_at  9.452307
## 321                   200731_s_at  7.536995
## 322                   200732_s_at  8.365855
## 323                   200733_s_at  9.763789
## 324                   200734_s_at  9.659388
## 325                   200735_x_at 14.044634
## 326                   200736_s_at 12.204499
## 327                     200737_at 11.190455
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## 823                     201233_at  9.985615
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## 1831                    202241_at  7.684442
## 1832                    202242_at  7.398639
## 1833                  202243_s_at 12.373394
## 1834                    202244_at 13.296083
## 1835                    202245_at  9.103868
## 1836                  202246_s_at 12.223053
## 1837                  202247_s_at  5.744794
## 1838                    202248_at  7.893126
## 1839                  202249_s_at  7.395488
## 1840                  202250_s_at  7.737834
## 1841                    202251_at  8.314418
## 1842                    202252_at 11.324235
## 1843                  202253_s_at  7.008488
## 1844                    202254_at  6.594934
## 1845                  202255_s_at  7.713812
## 1846                    202256_at  8.631466
## 1847                  202257_s_at  9.282025
## 1848                  202258_s_at  8.796846
## 1849                  202259_s_at  7.240132
## 1850                  202260_s_at  7.936116
## 1851                    202261_at  9.117536
## 1852                  202262_x_at  6.958926
## 1853                    202263_at  9.262282
## 1854                  202264_s_at  8.315849
## 1855                    202265_at 11.251478
## 1856                    202266_at 10.887666
## 1857                    202267_at  4.598880
## 1858                  202268_s_at 12.130947
## 1859                  202269_x_at  5.402992
## 1860                    202270_at  4.713239
## 1861                    202271_at  8.573513
## 1862                  202272_s_at 10.786676
## 1863                    202273_at  5.346634
## 1864                    202274_at  5.221612
## 1865                    202275_at  6.499682
## 1866                    202276_at 10.392043
## 1867                    202277_at 11.222435
## 1868                  202278_s_at  6.890296
## 1869                    202279_at 11.421441
## 1870                    202280_at  4.198843
## 1871                    202281_at  7.660665
## 1872                    202282_at 12.057405
## 1873                    202283_at  9.005760
## 1874                  202284_s_at  8.975238
## 1875                  202285_s_at  4.949499
## 1876                  202286_s_at  3.215502
## 1877                  202287_s_at  3.671672
## 1878                    202288_at  7.746435
## 1879                  202289_s_at  5.888200
## 1880                    202290_at  9.970313
## 1881                  202291_s_at  3.467102
## 1882                  202292_x_at  8.893630
## 1883                    202293_at  7.903109
## 1884                    202294_at  8.680664
## 1885                  202295_s_at  9.708879
## 1886                  202296_s_at 11.896650
## 1887                  202297_s_at 11.255384
## 1888                    202298_at 12.261516
## 1889                  202299_s_at 11.043190
## 1890                    202300_at 10.994053
## 1891                  202301_s_at  9.230291
## 1892                  202302_s_at 10.601847
## 1893                  202303_x_at  9.914580
## 1894                    202304_at  6.214587
## 1895                  202305_s_at  9.152770
## 1896                    202306_at 12.316978
## 1897                  202307_s_at 11.156130
## 1898                    202308_at  7.190552
## 1899                    202309_at 11.631239
## 1900                  202310_s_at  6.071148
## 1901                  202311_s_at  4.199248
## 1902                  202312_s_at  5.809587
## 1903                    202313_at 10.034328
## 1904                    202314_at  8.656592
## 1905                  202315_s_at  8.564067
## 1906                  202316_x_at  6.103482
## 1907                  202317_s_at  8.606294
## 1908                  202318_s_at  9.191359
## 1909                    202319_at  6.219870
## 1910                    202320_at  7.722578
## 1911                    202321_at  5.961478
## 1912                  202322_s_at  8.289641
## 1913                  202323_s_at  8.371504
## 1914                  202324_s_at 10.431373
## 1915                  202325_s_at 12.993725
## 1916                    202326_at  7.741717
## 1917                  202327_s_at  4.707999
## 1918                  202328_s_at  5.863950
## 1919                    202329_at 10.467019
## 1920                  202330_s_at 10.958263
## 1921                    202331_at  7.874252
## 1922                    202332_at  7.992211
## 1923                  202333_s_at 10.634706
## 1924                  202334_s_at  8.516983
## 1925                  202335_s_at  4.305595
## 1926                  202336_s_at  9.231889
## 1927                    202337_at  7.746113
## 1928                    202338_at  8.674722
## 1929                    202339_at  6.936373
## 1930                  202340_x_at  6.238685
## 1931                  202341_s_at  7.214152
## 1932                  202342_s_at  7.146408
## 1933                  202343_x_at 11.204780
## 1934                    202344_at  8.023976
## 1935                  202345_s_at 12.362422
## 1936                    202346_at  8.375386
## 1937                  202347_s_at 10.041433
## 1938                  202348_s_at  9.385348
## 1939                    202349_at  9.666213
## 1940                  202350_s_at  3.952312
## 1941                    202351_at  8.270475
## 1942                  202352_s_at 11.082215
## 1943                  202353_s_at  9.318388
## 1944                  202354_s_at  7.633184
## 1945                  202355_s_at  7.656160
## 1946                  202356_s_at  8.028861
## 1947                  202357_s_at  8.930687
## 1948                  202358_s_at  7.430162
## 1949                  202359_s_at  7.329938
## 1950                    202360_at  9.644629
## 1951                    202361_at  9.505528
## 1952                    202362_at 10.008351
## 1953                    202363_at  3.850304
## 1954                    202364_at  9.276186
## 1955                    202365_at  9.966894
## 1956                    202366_at  6.964312
## 1957                    202367_at  6.751533
## 1958                  202368_s_at  6.854769
## 1959                  202369_s_at  8.900297
## 1960                  202370_s_at 11.721516
## 1961                    202371_at  3.401266
## 1962                    202372_at  8.720843
## 1963                  202373_s_at  9.836002
## 1964                  202374_s_at  8.235865
## 1965                    202375_at  7.069298
## 1966                    202376_at  4.676731
## 1967                    202377_at  9.012089
## 1968                  202378_s_at  9.128962
## 1969                  202379_s_at  9.021805
## 1970                  202380_s_at  7.733336
## 1971                    202381_at  8.572983
## 1972                  202382_s_at  8.647901
## 1973                    202383_at  6.689769
## 1974                  202384_s_at  8.853132
## 1975                  202385_s_at  8.581497
## 1976                  202386_s_at  8.515243
## 1977                    202387_at  9.700258
## 1978                    202388_at 10.516479
## 1979                  202389_s_at  7.252817
## 1980                  202390_s_at  7.313835
## 1981                    202391_at 11.078905
## 1982                  202392_s_at  6.859394
## 1983                  202393_s_at  9.804249
## 1984                  202394_s_at  7.592661
## 1985                    202395_at  8.637469
## 1986                    202396_at 10.818126
## 1987                    202397_at 12.976520
## 1988                    202398_at  6.076979
## 1989                  202399_s_at  9.334168
## 1990                  202400_s_at  4.626039
## 1991                  202401_s_at  7.543658
## 1992                  202402_s_at  9.576485
## 1993                  202403_s_at  4.364852
## 1994                  202404_s_at  3.382286
## 1995                    202405_at  7.765531
## 1996                  202406_s_at 10.726833
## 1997                  202407_s_at  7.888099
## 1998                  202408_s_at 10.425935
## 1999                    202409_at  5.506329
## 2000                  202410_x_at  5.006357
## 2001                    202411_at  5.395439
## 2002                  202412_s_at 10.499580
## 2003                  202413_s_at 12.072147
## 2004                    202414_at  8.226676
## 2005                  202415_s_at  8.303930
## 2006                    202416_at 10.848895
## 2007                    202417_at  9.898027
## 2008                    202418_at  9.288490
## 2009                    202419_at  9.521736
## 2010                  202420_s_at 10.895146
## 2011                    202421_at  4.644919
## 2012                  202422_s_at  7.879945
## 2013                    202423_at  9.086154
## 2014                    202424_at  8.709438
## 2015                  202425_x_at  9.184636
## 2016                  202426_s_at  7.355123
## 2017                  202427_s_at 11.038221
## 2018                  202428_x_at 12.645470
## 2019                  202429_s_at  9.496508
## 2020                  202430_s_at  6.227929
## 2021                  202431_s_at 10.411344
## 2022                    202432_at  9.508685
## 2023                    202433_at 10.002536
## 2024                  202434_s_at  4.995847
## 2025                  202435_s_at 10.774250
## 2026                  202436_s_at 11.263980
## 2027                  202437_s_at 11.008938
## 2028                  202438_x_at  5.466779
## 2029                  202439_s_at  7.683158
## 2030                  202440_s_at  5.054074
## 2031                    202441_at 10.354388
## 2032                    202442_at 12.125699
## 2033                  202443_x_at 10.189548
## 2034                  202444_s_at  6.440116
## 2035                  202445_s_at  6.197914
## 2036                  202446_s_at  8.104274
## 2037                    202447_at  9.468880
## 2038                  202448_s_at  4.611597
## 2039                  202449_s_at  8.483527
## 2040                  202450_s_at  4.798054
## 2041                    202451_at  9.666396
## 2042                    202452_at  6.287903
## 2043                  202453_s_at  7.910575
## 2044                  202454_s_at  3.206858
## 2045                    202455_at  6.722614
## 2046                  202456_s_at  6.024025
## 2047                  202457_s_at  9.831284
## 2048                    202458_at  4.140238
## 2049                  202459_s_at  6.848527
## 2050                  202460_s_at  7.724215
## 2051                    202461_at 11.020007
## 2052                  202462_s_at 10.813285
## 2053                  202463_s_at  7.413009
## 2054                  202464_s_at  7.420224
## 2055                    202465_at  6.194514
## 2056                    202466_at  8.668968
## 2057                  202467_s_at 11.577852
## 2058                  202468_s_at  8.377804
## 2059                  202469_s_at 11.011384
## 2060                  202470_s_at  8.081099
## 2061                  202471_s_at  8.325032
## 2062                    202472_at  7.093231
## 2063                  202473_x_at  5.624525
## 2064                  202474_s_at  9.205529
## 2065                    202475_at 10.186615
## 2066                  202476_s_at  5.790160
## 2067                  202477_s_at  7.644204
## 2068                    202478_at  7.294952
## 2069                  202479_s_at  5.990202
## 2070                  202480_s_at  6.717571
## 2071                    202481_at  4.726522
## 2072                  202482_x_at  7.216756
## 2073                  202483_s_at 12.455390
## 2074                  202484_s_at 10.371614
## 2075                  202485_s_at  4.241970
## 2076                    202486_at  8.964437
## 2077                  202487_s_at 11.592465
## 2078                  202488_s_at  5.837736
## 2079                  202489_s_at  5.449561
## 2080                    202490_at  4.138447
## 2081                  202491_s_at 10.089221
## 2082                    202492_at  6.946459
## 2083                  202493_x_at  4.056515
## 2084                    202494_at  8.145953
## 2085                    202495_at  8.065551
## 2086                    202496_at  9.267304
## 2087                  202497_x_at  8.112056
## 2088                  202498_s_at  7.183502
## 2089                  202499_s_at  8.487296
## 2090                    202500_at  6.845734
## 2091                    202501_at  7.810213
## 2092                    202502_at 10.240131
## 2093                  202503_s_at 12.741390
## 2094                    202504_at  5.151056
## 2095                    202505_at 11.160949
## 2096                    202506_at  8.819579
## 2097                  202507_s_at  5.868045
## 2098                  202508_s_at  6.569701
## 2099                  202509_s_at  4.685756
## 2100                  202510_s_at  5.285491
## 2101                  202511_s_at  9.651518
## 2102                  202512_s_at  6.554541
## 2103                  202513_s_at  7.844252
## 2104                    202514_at  8.487349
## 2105                    202515_at  9.084033
## 2106                  202516_s_at  5.857884
## 2107                    202517_at  5.635387
## 2108                    202518_at  7.053210
## 2109                    202519_at  8.325870
## 2110                  202520_s_at 10.580508
## 2111                    202521_at 10.726666
## 2112                    202522_at 10.563210
## 2113                  202523_s_at  5.399244
## 2114                  202524_s_at  5.420247
## 2115                    202525_at  4.597815
## 2116                    202526_at  3.482364
## 2117                  202527_s_at  8.644765
## 2118                    202528_at  7.604335
## 2119                    202529_at  9.714560
## 2120                    202530_at  8.882754
## 2121                    202531_at  8.783974
## 2122                  202532_s_at 10.229012
## 2123                  202533_s_at  9.086659
## 2124                  202534_x_at 10.893641
## 2125                    202535_at  8.620967
## 2126                    202536_at  9.503951
## 2127                  202537_s_at  8.934826
## 2128                  202538_s_at  8.822202
## 2129                  202539_s_at  9.751501
## 2130                  202540_s_at  9.955105
## 2131                    202541_at 10.425124
## 2132                  202542_s_at 10.976809
## 2133                  202543_s_at  9.402375
## 2134                    202544_at 10.756631
## 2135                    202545_at  7.599598
## 2136                    202546_at  9.955920
## 2137                  202547_s_at  6.523392
## 2138                  202548_s_at  8.600740
## 2139                    202549_at  5.217872
## 2140                  202550_s_at  9.248019
## 2141                  202551_s_at  8.200980
## 2142                  202552_s_at  7.489896
## 2143                  202553_s_at  9.796730
## 2144                  202554_s_at  4.246476
## 2145                  202555_s_at  4.582535
## 2146                  202556_s_at  8.053463
## 2147                    202557_at 10.046807
## 2148                  202558_s_at  7.090221
## 2149                  202559_x_at  9.490023
## 2150                  202560_s_at 10.557472
## 2151                    202561_at  7.989050
## 2152                  202562_s_at 10.208908
## 2153                    202563_at  4.822106
## 2154                  202564_x_at  9.311650
## 2155                  202565_s_at  7.695323
## 2156                  202566_s_at  6.052798
## 2157                    202567_at 12.512926
## 2158                  202568_s_at  9.459217
## 2159                  202569_s_at  9.514213
## 2160                  202570_s_at  5.221289
## 2161                  202571_s_at  6.008070
## 2162                  202572_s_at  4.637649
## 2163                    202573_at  9.576552
## 2164                  202574_s_at  8.468951
## 2165                    202575_at  6.653492
## 2166                  202576_s_at  9.750260
## 2167                  202577_s_at 11.345769
## 2168                  202578_s_at  9.653832
## 2169                  202579_x_at 11.328264
## 2170                  202580_x_at  9.165993
## 2171                    202581_at 10.760039
## 2172                  202582_s_at 10.626799
## 2173                  202583_s_at  9.046784
## 2174                    202584_at  8.745194
## 2175                  202585_s_at  7.320822
## 2176                    202586_at  7.279416
## 2177                  202587_s_at  5.401925
## 2178                    202588_at  5.947804
## 2179                    202589_at 13.008096
## 2180                  202590_s_at  6.064832
## 2181                  202591_s_at 13.333268
## 2182                    202592_at  8.519323
## 2183                  202593_s_at  9.342917
## 2184                    202594_at  8.760559
## 2185                  202595_s_at  9.369880
## 2186                    202596_at 10.621584
## 2187                    202597_at  5.432419
## 2188                    202598_at  9.460379
## 2189                  202599_s_at  7.892008
## 2190                  202600_s_at  7.044132
## 2191                  202601_s_at  7.279586
## 2192                  202602_s_at 10.696782
## 2193                    202603_at 11.100120
## 2194                  202604_x_at  7.253644
## 2195                    202605_at 10.362875
## 2196                  202606_s_at 10.665770
## 2197                    202607_at  6.587700
## 2198                  202608_s_at  5.254878
## 2199                    202609_at  5.137669
## 2200                  202610_s_at  9.494984
## 2201                  202611_s_at  8.053941
## 2202                  202612_s_at  3.314831
## 2203                    202613_at  9.265553
## 2204                    202614_at  9.252305
## 2205                    202615_at  3.660202
## 2206                  202616_s_at  7.296939
## 2207                  202617_s_at  6.320500
## 2208                  202618_s_at  6.403803
## 2209                  202619_s_at  7.901483
## 2210                  202620_s_at  8.435899
## 2211                    202621_at  6.740703
## 2212                  202622_s_at  8.291762
## 2213                    202623_at 10.037001
## 2214                  202624_s_at  7.154515
## 2215                    202625_at  9.509706
## 2216                  202626_s_at  9.980446
## 2217                  202627_s_at  5.251831
## 2218                  202628_s_at  4.645993
## 2219                    202629_at  8.158141
## 2220                    202630_at  7.477195
## 2221                  202631_s_at  7.779644
## 2222                    202632_at  8.413928
## 2223                    202633_at 10.571131
## 2224                    202634_at  9.443906
## 2225                  202635_s_at 10.845397
## 2226                    202636_at  8.902914
## 2227                  202637_s_at 10.669106
## 2228                  202638_s_at 10.605653
## 2229                  202639_s_at  7.045544
## 2230                  202640_s_at  8.072350
## 2231                    202641_at  9.184308
## 2232                  202642_s_at  9.161127
## 2233                  202643_s_at  8.448274
## 2234                  202644_s_at  9.664377
## 2235                  202645_s_at  8.269775
## 2236                  202646_s_at  9.667082
## 2237                  202647_s_at  9.538950
## 2238                    202648_at  7.178660
## 2239                  202649_x_at 14.359197
## 2240                  202650_s_at  6.580880
## 2241                    202651_at 10.891687
## 2242                    202652_at  5.531156
## 2243                  202653_s_at 10.145183
## 2244                  202654_x_at  9.597731
## 2245                    202655_at 11.636639
## 2246                  202656_s_at  9.903499
## 2247                  202657_s_at 10.142828
## 2248                    202658_at  8.934457
## 2249                    202659_at 11.671374
## 2250                    202660_at  6.482243
## 2251                    202661_at  4.712864
## 2252                  202662_s_at  6.252733
## 2253                    202663_at  8.303122
## 2254                    202664_at  9.981229
## 2255                  202665_s_at  7.336433
## 2256                  202666_s_at 10.440014
## 2257                  202667_s_at  8.803779
## 2258                    202668_at  8.344622
## 2259                  202669_s_at  6.328883
## 2260                    202670_at 10.144633
## 2261                  202671_s_at  9.147329
## 2262                  202672_s_at 10.565800
## 2263                    202673_at 11.525233
## 2264                  202674_s_at  3.189083
## 2265                    202675_at 10.972273
## 2266                  202676_x_at  9.064727
## 2267                    202677_at  9.318142
## 2268                    202678_at 12.079049
## 2269                    202679_at  8.579126
## 2270                    202680_at  8.737434
## 2271                    202681_at  8.078452
## 2272                  202682_s_at  9.076212
## 2273                  202683_s_at  8.775698
## 2274                  202684_s_at  6.498409
## 2275                  202685_s_at  4.851707
## 2276                  202686_s_at  4.485458
## 2277                  202687_s_at  9.861029
## 2278                    202688_at  9.865781
## 2279                    202689_at  8.725440
## 2280                  202690_s_at 12.178454
## 2281                    202691_at  9.272548
## 2282                  202692_s_at  7.104959
## 2283                  202693_s_at  8.563498
## 2284                    202694_at  4.620242
## 2285                  202695_s_at  6.058297
## 2286                    202696_at 10.170783
## 2287                    202697_at 10.059398
## 2288                  202698_x_at 13.485129
## 2289                  202699_s_at  5.335148
## 2290                  202700_s_at  5.604524
## 2291                    202701_at  6.167203
## 2292                    202702_at  7.882871
## 2293                    202703_at 10.635639
## 2294                    202704_at  9.856038
## 2295                    202705_at 11.721956
## 2296                  202706_s_at  8.343322
## 2297                    202707_at  4.663833
## 2298                  202708_s_at  5.572704
## 2299                    202709_at  5.127695
## 2300                    202710_at  9.051432
## 2301                    202711_at  5.728353
## 2302                  202712_s_at  4.844824
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## 10305                 210783_x_at  5.582946
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## 10532                 211024_s_at  4.353125
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## 10536                 211028_s_at  8.339181
## 10537                 211029_x_at  4.330691
## 10538                 211030_s_at  5.321074
## 10539                 211031_s_at  7.189312
## 10540                   211032_at  3.607518
## 10541                 211033_s_at  5.924345
## 10542                 211034_s_at  7.707851
## 10543                 211036_x_at 11.914660
## 10544                 211037_s_at  5.994012
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## 10547                 211040_x_at  9.737666
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## 10553                 211047_x_at 11.022260
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## 10555                   211049_at  6.663909
## 10556                 211050_x_at  5.256233
## 10557                 211051_s_at  5.371543
## 10558                 211052_s_at  8.067200
## 10559                   211053_at  4.448874
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## 10561                 211055_s_at  4.434691
## 10562                 211056_s_at  7.132239
## 10563                   211057_at  4.738693
## 10564                 211058_x_at 13.788560
## 10565                 211059_s_at  6.832012
## 10566                 211060_x_at  8.170856
## 10567                 211061_s_at  9.671070
## 10568                 211062_s_at  5.541944
## 10569                 211063_s_at  9.604846
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## 10571                 211065_x_at  8.228541
## 10572                 211066_x_at  4.876001
## 10573                 211067_s_at  6.890190
## 10574                 211068_x_at  7.095181
## 10575                 211069_s_at 11.527163
## 10576                 211070_x_at 12.990235
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## 10582                 211076_x_at  5.772427
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## 10586                 211080_s_at  6.658628
## 10587                 211081_s_at  7.172227
## 10588                 211082_x_at  4.648263
## 10589                 211083_s_at  3.611110
## 10590                 211084_x_at  7.894110
## 10591                 211085_s_at  7.726023
## 10592                 211086_x_at  5.722797
## 10593                 211087_x_at  5.278424
## 10594                 211088_s_at  4.406915
## 10595                 211089_s_at  6.779223
## 10596                 211090_s_at  7.913690
## 10597                 211091_s_at  5.339554
## 10598                 211092_s_at  3.965951
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## 10603                 211097_s_at  5.506230
## 10604                 211098_x_at  9.433542
## 10605                 211099_s_at  4.606538
## 10606                 211100_x_at  5.016762
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## 10608                 211102_s_at  4.222580
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## 10624                 211118_x_at  4.606452
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## 10636                 211130_x_at  5.202550
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## 10647                 211142_x_at  5.997689
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## 10692                 211189_x_at  4.477566
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## 10704                   211201_at  4.485795
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## 10707                   211204_at  4.452013
## 10708                 211205_x_at  5.930407
## 10709                 211207_s_at  3.024564
## 10710                 211208_s_at  7.997724
## 10711                 211209_x_at  7.371981
## 10712                 211210_x_at  8.069751
## 10713                 211211_x_at  7.936178
## 10714                 211212_s_at  7.412432
## 10715                   211213_at  3.800087
## 10716                 211214_s_at  4.791553
## 10717                 211215_x_at  5.741401
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## 10719                   211218_at  3.101426
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## 10722                   211221_at  4.246981
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## 10727                   211226_at  4.409915
## 10728                 211227_s_at  3.037987
## 10729                 211228_s_at  7.782928
## 10730                 211230_s_at  5.501215
## 10731                 211231_x_at  5.512099
## 10732                 211232_x_at  4.825021
## 10733                 211233_x_at  6.162835
## 10734                 211234_x_at  6.067781
## 10735                 211235_s_at  5.565494
## 10736                 211237_s_at  5.382522
## 10737                   211238_at  4.668965
## 10738                 211239_s_at  2.708044
## 10739                 211240_x_at  7.383500
## 10740                   211241_at  4.749180
## 10741                 211242_x_at  4.602463
## 10742                 211245_x_at  4.245953
## 10743                 211248_s_at  5.602840
## 10744                   211249_at  5.424270
## 10745                 211250_s_at  5.441732
## 10746                 211251_x_at  8.013455
## 10747                 211252_x_at  7.157038
## 10748                 211253_x_at  5.948207
## 10749                 211254_x_at  4.973819
## 10750                 211255_x_at  6.602989
## 10751                 211256_x_at  7.133100
## 10752                 211257_x_at  9.743269
## 10753                 211258_s_at  4.519272
## 10754                 211259_s_at  6.101994
## 10755                   211260_at  5.884410
## 10756                   211261_at  5.991590
## 10757                   211262_at  4.265597
## 10758                 211263_s_at  5.708922
## 10759                   211264_at  3.034609
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## 10761                 211266_s_at  5.281315
## 10762                   211267_at  5.326087
## 10763                 211269_s_at  5.303317
## 10764                 211270_x_at 12.145892
## 10765                 211271_x_at 11.070756
## 10766                 211272_s_at  4.503381
## 10767                 211273_s_at  3.208172
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## 10769                 211275_s_at 10.480233
## 10770                   211276_at  3.985608
## 10771                 211277_x_at  5.810741
## 10772                   211279_at  5.410989
## 10773                 211280_s_at  5.469485
## 10774                 211282_x_at  6.041108
## 10775                 211284_s_at  8.570994
## 10776                 211285_s_at 10.630767
## 10777                 211286_x_at  4.212558
## 10778                 211287_x_at  4.084031
## 10779                 211289_x_at  8.111411
## 10780                 211295_x_at  5.353259
## 10781                 211296_x_at 13.806259
## 10782                 211297_s_at  9.703525
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## 10850                 211372_s_at 10.979490
## 10851                 211373_s_at  8.218863
## 10852                 211374_x_at  5.824208
## 10853                 211375_s_at 10.758965
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## 10856                 211378_x_at 13.377827
## 10857                 211379_x_at  2.388063
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## 10863                 211385_x_at  6.843146
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## 10866                 211388_s_at  4.536128
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## 10887                   211412_at  3.022276
## 10888                 211413_s_at  4.570509
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## 10891                 211417_x_at  8.534745
## 10892                 211419_s_at  5.806127
## 10893                 211421_s_at  2.985654
## 10894                   211422_at  3.736433
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## 10898                 211426_x_at  6.360709
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## 10902                 211430_s_at  4.585928
## 10903                 211431_s_at  5.358302
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## 10995                 211527_x_at  4.788185
## 10996                 211528_x_at 10.046172
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## 10998                 211530_x_at  9.293127
## 10999                 211531_x_at  6.538336
## 11000                 211532_x_at  4.241231
## 11001                   211533_at  2.831964
## 11002                 211534_x_at  8.001653
## 11003                 211535_s_at  8.326600
## 11004                 211536_x_at  7.366754
## 11005                 211537_x_at  8.131501
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## 11010                 211543_s_at  6.915596
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## 11076                 211609_x_at 11.291702
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## 11090                 211623_s_at 12.092597
## 11091                 211624_s_at  4.949272
## 11092                 211625_s_at  3.621910
## 11093                 211626_x_at  4.933809
## 11094                 211627_x_at  3.740896
## 11095                 211628_x_at 12.734161
## 11096                 211630_s_at  8.161382
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## 11099                 211633_x_at  4.703917
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## 11192                 211730_s_at 12.088085
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## 11194                 211732_x_at  6.754924
## 11195                 211733_x_at 11.056321
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## 11317                 211866_x_at  5.323122
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## 11325                 211874_s_at  5.966718
## 11326                 211875_x_at  4.580056
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## 49844                1559856_s_at  3.435539
## 49845                  1559861_at  5.581755
## 49846                  1559862_at  3.431561
## 49847                1559863_a_at  5.127747
## 49848                  1559864_at  5.084541
## 49849                  1559865_at  3.105431
## 49850                  1559867_at  5.523243
## 49851                1561114_a_at  5.219589
## 49852                  1559870_at  5.575242
## 49853                1559871_s_at  3.829872
## 49854                  1559876_at  5.019572
## 49855                  1559877_at  4.280188
## 49856                1559879_a_at  3.276485
## 49857                  1559880_at  4.426519
## 49858                1559881_s_at  6.820563
## 49859                  1559882_at  2.951280
## 49860                1559883_s_at  3.938526
## 49861                  1559884_at  3.322261
## 49862                  1559889_at  4.202337
## 49863                1559890_a_at  2.844113
## 49864                  1559891_at  4.606163
## 49865                  1559893_at  5.056769
## 49866                1559895_x_at  3.717335
## 49867                1565228_s_at  2.852801
## 49868                1559901_s_at  4.972292
## 49869                  1559902_at  4.783412
## 49870                1559909_a_at  5.120009
## 49871                  1559910_at  4.902276
## 49872                  1559911_at  3.176326
## 49873                  1559914_at  2.882754
## 49874                  1559915_at  4.686586
## 49875                1559916_a_at  4.794245
## 49876                1559917_a_at  5.524846
## 49877                1559920_a_at  3.931158
## 49878                  1559921_at  5.566782
## 49879                  1559922_at  5.041825
## 49880                  1559923_at  3.001290
## 49881                  1559924_at  5.317780
## 49882                1559925_s_at  4.124320
## 49883                  1559926_at  5.876473
## 49884                1559927_a_at  5.291028
## 49885                  1559928_at  3.802749
## 49886                  1559929_at  3.918801
## 49887                  1559930_at  2.310068
## 49888                  1559932_at  3.709285
## 49889                  1559936_at  3.601904
## 49890                  1559939_at  3.469009
## 49891                1559940_s_at  4.915819
## 49892                  1559942_at  8.283576
## 49893                  1559945_at  4.571441
## 49894                1559946_s_at 12.060021
## 49895                  1559948_at  4.300923
## 49896                  1559949_at  2.503238
## 49897                  1559950_at  3.724799
## 49898                1559952_x_at  4.146176
## 49899                  1559953_at  3.408829
## 49900                1559954_s_at  6.570908
## 49901                  1559956_at  4.448308
## 49902                1559957_a_at  6.625603
## 49903                1559960_x_at  4.554741
## 49904                  1569867_at  3.868214
## 49905                1569868_s_at  6.014437
## 49906                  1559964_at  3.544529
## 49907                  1559965_at  2.832301
## 49908                1559966_a_at  3.412940
## 49909                  1559970_at  2.903010
## 49910                  1559971_at  4.008362
## 49911                1560779_a_at  3.242028
## 49912                  1559975_at  4.890266
## 49913                  1559976_at  4.154912
## 49914                1559977_a_at  5.194870
## 49915                  1559979_at  4.236371
## 49916                  1559980_at  4.034882
## 49917                1559982_s_at  4.284922
## 49918                  1559986_at  2.577737
## 49919                  1559987_at  3.885862
## 49920                  1559988_at  4.143478
## 49921                  1559989_at  5.599989
## 49922                  1559990_at  3.142747
## 49923                1559991_s_at  4.583763
## 49924                1559992_a_at  2.386535
## 49925                  1559993_at  7.284075
## 49926                  1559995_at  5.448935
## 49927                1559996_s_at  5.391509
## 49928                1559997_x_at  5.300716
## 49929                  1560001_at  4.840550
## 49930                  1560002_at  4.735466
## 49931                1560006_a_at  5.272466
## 49932                  1560007_at  4.039347
## 49933                1560010_a_at  4.217594
## 49934                  1560011_at  4.613690
## 49935                  1560012_at  3.148340
## 49936                  1560013_at  5.157034
## 49937                1560014_s_at  5.256121
## 49938                  1560017_at  5.966949
## 49939                  1560018_at  2.869174
## 49940                  1560019_at  3.404327
## 49941                  1560020_at  4.195645
## 49942                  1560021_at  4.345997
## 49943                1560023_x_at  4.911993
## 49944                  1560024_at  4.369460
## 49945                  1560025_at  4.840281
## 49946                  1560026_at  4.451138
## 49947                  1560028_at  3.591151
## 49948                1560029_a_at  3.481976
## 49949                  1560030_at  3.894180
## 49950                  1560031_at  3.860081
## 49951                  1560033_at  4.030683
## 49952                1560034_a_at  3.343461
## 49953                  1560035_at  5.199617
## 49954                  1560038_at  3.214297
## 49955                  1560042_at  4.226645
## 49956                  1560043_at  4.126209
## 49957                1560047_s_at  4.874019
## 49958                  1560048_at  3.231632
## 49959                  1560049_at  4.679500
## 49960                  1560051_at  3.058721
## 49961                  1560058_at  4.874788
## 49962                  1560059_at  5.838865
## 49963                1560060_s_at  9.751424
## 49964                  1560062_at  4.981271
## 49965                1560063_a_at  3.182951
## 49966                  1560065_at  4.251447
## 49967                  1560066_at  5.158509
## 49968                1560068_a_at  4.347552
## 49969                  1560069_at  4.230301
## 49970                  1560070_at  3.961065
## 49971                1560071_a_at  5.658801
## 49972                  1560074_at  4.915658
## 49973                  1560075_at  3.222060
## 49974                  1560078_at  3.854081
## 49975                  1560080_at  3.110333
## 49976                  1560081_at  5.299379
## 49977                  1560082_at  4.676392
## 49978                  1560086_at  2.942020
## 49979                1560087_a_at  3.834042
## 49980                  1560089_at  6.600788
## 49981                  1560094_at  4.548987
## 49982                1560095_s_at  4.667635
## 49983                  1560097_at  5.053717
## 49984                  1560098_at  2.948821
## 49985                  1560099_at  6.904948
## 49986                  1560100_at  5.560983
## 49987                  1560101_at  4.914911
## 49988                  1560102_at  4.518731
## 49989                  1560104_at  3.193353
## 49990                  1560105_at  3.620768
## 49991                  1560107_at  2.619142
## 49992                  1560108_at  3.911528
## 49993                1560109_s_at  4.385759
## 49994                  1560111_at  2.472636
## 49995                  1560112_at  3.170540
## 49996                1560115_a_at  5.695858
## 49997                1560116_a_at 10.109542
## 49998                  1560117_at  4.449395
## 49999                  1560118_at  3.519767
##  [ reached 'max' / getOption("max.print") -- omitted 4676 rows ]

GSM - Sample

pryr::object_size(gsm)
## 4.39 MB
class(gsm)
## [1] "GSM"
## attr(,"package")
## [1] "GEOquery"
isS4(gsm)
## [1] TRUE

GPL - Platform

Here is obtaining GPL metadata.

gpl <- getGEO("GPL570", destdir = "GeoData")
pryr::object_size(gpl)
## 74.01 MB
class(gpl)
## [1] "GPL"
## attr(,"package")
## [1] "GEOquery"

GPL - Platform

Meta(gpl) %>%
    str(vec.len = 1, nchar.max = 65)
## List of 27
##  $ contact_address        : chr " "
##  $ contact_city           : chr "Santa Clara"
##  $ contact_country        : chr "USA"
##  $ contact_email          : chr "geo@ncbi.nlm.nih.gov, support@affymetrix.com"
##  $ contact_institute      : chr "Affymetrix, Inc."
##  $ contact_name           : chr " ,,Affymetrix, Inc."
##  $ contact_phone          : chr "888-362-2447"
##  $ contact_state          : chr "CA"
##  $ contact_web_link       : chr "http://www.affymetrix.com/index.affx"
##  $ contact_zip/postal_code: chr "95051"
##  $ data_row_count         : chr "54675"
##  $ description            : chr [1:5] "Affymetrix submissions are typically submitted t"| __truncated__ ...
##  $ distribution           : chr "commercial"
##  $ geo_accession          : chr "GPL570"
##  $ last_update_date       : chr "Dec 14 2020"
##  $ manufacture_protocol   : chr [1:6] "see manufacturer's web site" ...
##  $ manufacturer           : chr "Affymetrix"
##  $ organism               : chr "Homo sapiens"
##  $ relation               : chr [1:61] "Alternative to: GPL4454 (Alternative CDF)" ...
##  $ sample_id              : chr [1:166727] "GSM18422" ...
##  $ series_id              : chr [1:5550] "GSE1145" ...
##  $ status                 : chr "Public on Nov 07 2003"
##  $ submission_date        : chr "Nov 07 2003"
##  $ taxid                  : chr "9606"
##  $ technology             : chr "in situ oligonucleotide"
##  $ title                  : chr "[HG-U133_Plus_2] Affymetrix Human Genome U133 Plus 2.0 Array"
##  $ web_link               : chr [1:2] "http://www.affymetrix.com/support/technical/bypr"| __truncated__ ...

GPL - Platform

Columns(gpl) %>%
    as_tibble()
## # A tibble: 16 × 2
##    Column                           Description                                 
##    <chr>                            <chr>                                       
##  1 ID                               "Affymetrix Probe Set ID LINK_PRE:\"https:/…
##  2 GB_ACC                           "GenBank Accession Number LINK_PRE:\"http:/…
##  3 SPOT_ID                          "identifies controls"                       
##  4 Species Scientific Name          "The genus and species of the organism repr…
##  5 Annotation Date                  "The date that the annotations for this pro…
##  6 Sequence Type                    ""                                          
##  7 Sequence Source                  "The database from which the sequence used …
##  8 Target Description               ""                                          
##  9 Representative Public ID         "The accession number of a representative s…
## 10 Gene Title                       "Title of Gene represented by the probe set…
## 11 Gene Symbol                      "A gene symbol, when one is available (from…
## 12 ENTREZ_GENE_ID                   "Entrez Gene Database UID LINK_PRE:\"http:/…
## 13 RefSeq Transcript ID             "References to multiple sequences in RefSeq…
## 14 Gene Ontology Biological Process "Gene Ontology Consortium Biological Proces…
## 15 Gene Ontology Cellular Component "Gene Ontology Consortium Cellular Componen…
## 16 Gene Ontology Molecular Function "Gene Ontology Consortium Molecular Functio…

GPL - Platform

Table(gpl) %>%
    dim()
## [1] 54675    16
Table(gpl) %>%
    as_tibble() %>%
    dplyr::select(ID, GB_ACC, `Sequence Source`, `Target Description`)
## # A tibble: 54,675 × 4
##    ID        GB_ACC `Sequence Source`               `Target Description`        
##    <chr>     <chr>  <chr>                           <chr>                       
##  1 1007_s_at U48705 Affymetrix Proprietary Database U48705 /FEATURE=mRNA /DEFIN…
##  2 1053_at   M87338 GenBank                         M87338 /FEATURE= /DEFINITIO…
##  3 117_at    X51757 Affymetrix Proprietary Database X51757 /FEATURE=cds /DEFINI…
##  4 121_at    X69699 GenBank                         X69699 /FEATURE= /DEFINITIO…
##  5 1255_g_at L36861 Affymetrix Proprietary Database L36861 /FEATURE=expanded_cd…
##  6 1294_at   L13852 GenBank                         L13852 /FEATURE= /DEFINITIO…
##  7 1316_at   X55005 Affymetrix Proprietary Database X55005 /FEATURE=mRNA /DEFIN…
##  8 1320_at   X79510 Affymetrix Proprietary Database X79510 /FEATURE=cds /DEFINI…
##  9 1405_i_at M21121 GenBank                         M21121 /FEATURE= /DEFINITIO…
## 10 1431_at   J02843 Affymetrix Proprietary Database J02843 /FEATURE=cds /DEFINI…
## # … with 54,665 more rows

GSE - Series

gse <- getGEO(GEO = "GSE54157", destdir = "GeoData", GSEMatrix = FALSE)  # Note the GSEMatrix argument
gse_exprs <- getGEO(GEO = "GSE54157", destdir = "GeoData", GSEMatrix = TRUE)  # Note the GSEMatrix argument

GSE - Series

class(gse)
## [1] "GSE"
## attr(,"package")
## [1] "GEOquery"
Meta(gse)[c("title", "type", "platform_id", "summary", "supplementary_file")]
## $title
## [1] "Expression changes induced by PTPN1 knockdown in KM-H2 cells"
## 
## $type
## [1] "Expression profiling by array"
## 
## $platform_id
## [1] "GPL570"
## 
## $summary
## [1] "Gene expression data for shRNA PTPN1 knockdown vs. Non-silencing in the classical Hodgkin lymphoma-derived cell line KM-H2"
## 
## $supplementary_file
## [1] "ftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE54nnn/GSE54157/suppl/GSE54157_RAW.tar"

GSE - Series

We can extract out the GSM and GPL in the series.

GEOquery::GSMList(gse) %>%
    names()
## [1] "GSM1308975" "GSM1308976" "GSM1308977" "GSM1308978"
GEOquery::GPLList(gse) %>%
    names()
## [1] "GPL570"

GSE - Series

gse_exprs <- getGEO(GEO = "GSE54157", destdir = "geoData", GSEMatrix = TRUE)  # Note the GSEMatrix argument

GSE - Series

class(gse_exprs)
## [1] "list"
length(gse_exprs)
## [1] 1
pryr::object_size(gse_exprs)
## 63.81 MB
lapply(gse_exprs, class)
## $GSE54157_series_matrix.txt.gz
## [1] "ExpressionSet"
## attr(,"package")
## [1] "Biobase"

GSE - Series

Phenotype data can be extracted from the ExpressionSet

gse_GPL570 <- gse_exprs[[1]]
Biobase::pData(gse_GPL570) %>%
    as_tibble()

GSE - Series

In this case, the GEO series does not have pre-processed expression data. Instead, raw data can be accessed from the supplemental file links. Here we get two files. The full path to the download is saved as a rowname.

supp_info <- GEOquery::getGEOSuppFiles(GEO = "GSE147507")
rownames(supp_info)
## [1] "/__w/RU_ReproducibleR_abridged/RU_ReproducibleR_abridged/extdata/GSE147507/GSE147507_RawReadCounts_Ferret.tsv.gz"
## [2] "/__w/RU_ReproducibleR_abridged/RU_ReproducibleR_abridged/extdata/GSE147507/GSE147507_RawReadCounts_Human.tsv.gz"
basename(rownames(supp_info)[1])
## [1] "GSE147507_RawReadCounts_Ferret.tsv.gz"
basename(rownames(supp_info)[2])
## [1] "GSE147507_RawReadCounts_Human.tsv.gz"

Creating metadata using template: CEDAR Workbench


CEDAR Workbench

It is an online resource that makes it easy to collect and use metadata

It helps user to

create user-friendly, shareable forms for collecting metadata, with features like share your forms and metadata * link your questions (fields) and possible answers (values) to controlled terms * view metadata responses meeting your search criteria, in several forms * use the Workbench Desktop interface to manage your content * enable intelligent metadata suggestions in your template by using a field’s Suggestions tab

Create a CEDAR account

Webpage: cedar.metadatacenter.org

Click on the “Register” link to

  • Create an account

CEDAR template

Once registered,

  • Click on the Shared with Me link on the left side to find sample template
  • Try creating your own

CEDAR and R

For installation:

devtools::install_github("earnaud/cedarr", dependencies = TRUE)

For help:

help("API response", package = "cedarr")
# ?cedarr::`API response`

Contact

Any suggestions, comments, edits or questions (about content or the slides themselves) please reach out to our GitHub and raise an issue.